[Histonet] rhodamine auto-fluor. problemI

[RCHIOVETTI <@t> aol.com]

In a message dated 10/24/2003 3:01:17 PM US Mountain Standard Time, 
Bonnie.P.Whitaker <@t> uth.tmc.edu writes:

> I have a question for you guys:  A researcher here in OB/GYN is doing
>  rhodamine-labeled fluorescent work (he didn't say what antibody) on mouse
>  fallopian tube... he thought he was having a staining problem, but has
>  determined that his tissue is auto-fluorescing.  The tissue is frozen in 
OCT
>  and fixed in acetone/methanol.  What can he do to quench this?
>  
Hi Bonnie,

Is this a new project, or a recent problem with an ongoing project?  Is the 
autofluorescence in the red wavelengths, or is the investigator perhaps seeing 
a yellow or green color in the scope?  I'm wondering if the scope the 
investigator is using has been successfully used for rhodamine work before?

Fluorescence filter sets are usually constructed in one of two ways: They may 
have a probe-specific "bandpass" filter in them which will only pass the 
specific wavelength of the probe (rhodamine, in this case), or a "longpass" filter 
which will pass the signal from the probe, plus other wavelengths which are 
close to, but not exactly the same as, the probe.

One thing's for sure, if your colleague is looking for red fluorescence and 
is seeing a yellowish or greenish color through the cube, he may need to use a 
different filter set in the scope.  The manufacturer of the scope or the 
manufacturer of the filter set should be able to give some guidance here.

Hope this is of some help!

Bob Chiovetti
GTI Microsystems
Leica Regional Dealer
Desert Southwest Region USA